I had the same problem and Bob (the guy who made those
yeast vectors) sugggested the following.. I am pasting as it is..
if you need further clarifications, pls feel free to mail me directly.
So here are your options:
1) Grow on SC-uracil-glucose to mid-log, spin and wash cells, then add
YEP-4%g galactose and induce
for greater than 6 hours. This should be ample time to overcome glucose
repression and induction
of the GST fusion promoter.
2) Grow in SC-uracil-glucose (2%) to establish a starter culture. Use
this to start a culture in
YEP-Raffinose (2%) or YEP-glycerol/ethanol (2%/2%). At mid-log, simply add
galactose to the
culture to make the final concetration 4%. You will begin seeing induction
within minutes, but for
maximum yield (assuming no toxicity), induce for greater than 6 hours.
Sailesh Surapureddi Ph.D Communicators
Plan #12, Cell Biology Work +46-13-223917
Dept. of Biomedicine and Surgery Res +46-13-138839
H=E4lsouniveristetet Cel +46-70-4916570
S 581 85, Link=F6ping Fax +46-13-224149
Sweden. Email Saisu at mcb.liu.se
Sometimes mother nature does not like to be mimicked!!!.........
=2E.......... One of the many reasons I give to my boss when I fumble up!!!